Bermuda grass 'Tifway' dehydrin-L, encoding gene and probe thereof

A dehydrin and probe technology, applied in genetic engineering, plant genetic improvement, recombinant DNA technology, etc., can solve the problem of no bermudagrass dehydrin protein structure and other problems

Active Publication Date: 2015-11-11
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no literature report related to the structure of bermudagrass dehydrin protein and its coding gene sequence

Method used

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  • Bermuda grass 'Tifway' dehydrin-L, encoding gene and probe thereof
  • Bermuda grass 'Tifway' dehydrin-L, encoding gene and probe thereof
  • Bermuda grass 'Tifway' dehydrin-L, encoding gene and probe thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 , Bermudagrass 'Tifway' Dehydrin-L gene cloning

[0046] 1. Acquisition of plant material

[0047] Take bermudagrass 'Tifway' leaf tissue for RNA extraction;

[0048] 2. Extraction of RNA

[0049] Total RNA (Trizol: Invitrogen) was extracted with "RNApreppure Plant Total RNA Extraction Kit", the integrity of RNA was identified by formaldehyde denaturing gel electrophoresis, and then the purity and concentration of RNA were determined on a spectrophotometer (ThermoScientificNANODROP1000Spectrophotometer);

[0050] 3. Full-length cloning of genes

[0051] According to the EST sequence and protein function annotation results obtained from the isolation of the suppression and subtraction library (SSH) established during the drought stress of Bermudagrass 'Tifway', the core fragment of the Dehydrin-L gene of Bermudagrass 'Tifway' was obtained. Using the RACE method (SMARTer TM RACE cDNA Amplification Kit: Clonetech) for full-length cDNA cloning in three stage...

Embodiment 2

[0064] Example 2 , Sequence information and homology analysis of the 'Tifway' Dehydrin-L gene of Bermudagrass

[0065] The full-length open reading frame sequence of the bermudagrass 'Tifway' Dehydrin-L gene of the present invention is 543bp, and the detailed sequence is shown in the sequence shown in SEQ ID NO.3. The amino acid sequence of Bermudagrass 'Tifway' Dehydrin-L protein was deduced according to the sequence of the open reading frame, with a total of 180 amino acid residues, a molecular weight of 18.2kDa, and an isoelectric point (pI) of 8.78. The detailed sequence is shown in SEQ ID NO.4 sequence;

[0066] The open reading frame sequence of Bermudagrass 'Tifway' Dehydrin-L and the amino acid sequence of its encoded protein were carried out in Non-redundantGenBank+EMBL+DDBJ+PDB and Non-redundantGenBankCDStranslations+PDB+SwissProt+Superdate+PIR databases using BLAST program Nucleotide and protein homology search, it was found that it has 78% identity at the nucleo...

Embodiment 3

[0067] Example 3 , Expression difference of bermudagrass 'Tifway' Dehydrin-L gene in different stages of drought stress

[0068] 1. Material acquisition: samples were taken from leaves of Bermudagrass 'Tifway' materials at different stages of drought stress (0 day, 5 days, 10 days, 15 days). Wrap the samples with aluminum platinum paper and put them into liquid nitrogen immediately, and then transfer them to -80°C ultra-low temperature refrigerator for storage until use;

[0069] 2. RNA extraction: use RNApreppure plant total RNA extraction (Trizol: Invitrogen); extract the total RNA in different sample tissues of Bermudagrass 'Tifway';

[0070] 3. Determination of the integrity, purity and concentration of RNA: use ordinary agarose gel electrophoresis (gel concentration 1.2;% 0.5×TBE electrophoresis buffer; 150v, 15min) to detect integrity, and the maximum rRNA brightness in the electrophoresis band should be 1.5-2.0 times the brightness of the second rRNA, otherwise it me...

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Abstract

The invention relates to bermuda grass 'Tifway' dehydrin-L, an encoding gene and a probe thereof. The dehydrin-L is a protein represented as (a) or (b), wherein (a) is a protein composed of the amino acid sequence represented as the Seq ID No.4 and (b) is a protein which is a derivative from the protein (a), is obtained through substitution, deficiency or addition of one or more amino acids and has the bermuda grass 'Tifway' dehydrin-L activity. The invention also provides a nucleic acid sequence for encoding the proteins and the probe for detecting the nucleic acid. In the invention, adversity stress physiological response, such as drought resistance and the like, of the bermuda grass 'Tifway' are regulated through genetic engineering technologies, thereby improving the drought resistance of turfgrass. The invention provides theoretical evidence for molecular breeding and has a huge application value.

Description

technical field [0001] The present invention relates to an important protective protein dehydrin-L and its encoding gene and probe in the response process of bermudagrass 'Tifway' to drought stress, and specifically relates to a bermudagrass 'Tifway' dehydrin protein Dehydrin-L and its encoding Genes and Probes. Background technique [0002] Bermudagrass (Cynodondactylon) is a very important warm-season turfgrass. It is a perennial herb of the Poaceae family. It has well-developed rhizomes and stolons. Good color and other advantages, widely used in sports grounds, playgrounds, parks and soil-fixing slope protection lawns at home and abroad. Therefore, it is one of the most valuable and widely used grass species among warm-season turfgrasses. It enjoys the title of "the master grass species" of warm-season turfgrasses, and has great social, economic and ecological values. Bermudagrass has a very strong ability to resist drought. The annual evapotranspiration is less than 4...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12Q1/68C12N15/11
Inventor 周鹏安渊吕爱敏张荻李姣姣
Owner SHANGHAI JIAO TONG UNIV
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